Injured Liver Conditional Culture Medium Induces Differentiation Of Mouse Bone Marrow Mononuclear Cells Into A Hepatocyte Lineage In Vitro

Objective: To explore whether the bone marrow mononuclear cells, BM MNCs of mice can be induced into hepatocytes and the condition of differentiation in vitro. Methods: The injured liver from the mouse exposed to hepatotoxic reagents carbon tetrachloride (CCl₄) was cultured, and the medium was collected. BM MNCs were cultured in the medium containing this conditional culture solution of injured liver. The morphological changes of BM MNCs were observed by phase-contrast microscope. On day 1, 7, 14, 21, mRNA of albumin, AFP, CK18, CK19, HNF3β and TAT of BM MNCs were examined by reverse transcription polymerase chain reaction (RT-PCR), and the expressions of these genes were also detected by immunofluorescence assay, and glycogen by PAS (Periodic acid-Schiff reaction). Results: In groups induced by conditional culture solution after 21 days, BM MNCs showed characteristic of hepatocytes; On day 7, AFP and CK19 mRNA of MNCs could be detected by RT-PCR, and AFP increased on day 14 but directed on day 21; Albumin, HNF3β and CK18 mRNA could also be detected from day 14 to day 21; and TAT can be detected on day 21 by RT-PCR. Immunofluorescence assay for CK18, CK19, albumin and AFP showed positive staining reaction on day 21. Differentiated cells further demonstrated these cells also acquired functional characteristics of hepatocytes that they can store glycogen. Conclusion: BM MNCs of adult mice cultured in conditional culture solution of injured liver can differentiate into hepatocytes. The establishment of this induction system may play an important role in the study to the mechanism of the differentiation of BM MNCs, and suggest a new source of seed cells for hepatocyte transplantation.